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Significance Thermostable Cas13 orthologues will expand and unlock the power of CRISPR systems for synthetic biology and biotechnology applications. We identified and characterized a thermostable Cas13 orthologue from Thermoclostridium caenicola… Click to show full abstract
Significance Thermostable Cas13 orthologues will expand and unlock the power of CRISPR systems for synthetic biology and biotechnology applications. We identified and characterized a thermostable Cas13 orthologue from Thermoclostridium caenicola (TccCas13a), which exhibits robust cis and trans catalytic activities over a broad range of temperatures from 37 to 70 °C. To demonstrate the utility of the thermostable Cas13 enzymes in enabling biotechnological applications, we coupled reverse transcription–loop-mediated isothermal amplification to TccCas13a to develop a one-pot assay for virus detection. The developed assay exhibits key point-of-care (POC) features, enabling its use for the detection of different pathogens. This platform was clinically validated and is suitable for multiplexed detection. TccCas13a will enable advancements in POC diagnostics, transcriptome engineering, editing, and targeted therapies.
Journal Title: Proceedings of the National Academy of Sciences of the United States of America
Year Published: 2022
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